Figure 1: SCLIM observation of chloroplasts in a live WT P. patens protonemal cell.

Chloroplasts in P. patens protonema grown on agar medium in a glass-bottom dish were observed directly. (a–c) 3D images of WT P. patens chloroplasts observed using conventional confocal microscopy. (a) Differential interference contrast image, (b) Chl fluorescence image, and (c) Chl fluorescence image with the same scale as in (d,e). The 3D image was taken using five optical sections (x, y) sequentially acquired at 0.2 μm intervals (z = 1 μm) as previously described⁴¹. (d–g) 3D images of WT P. patens chloroplasts observed using SCLIM. (d) Chl fluorescence image before 3D deconvolution, (e) Chl fluorescence image after 3D deconvolution, and (f,g) close-up views of (e) from different angles. Arrows and arrowheads indicate the same structures in different figures (e–g). 3D image reconstructed using 100 optical sections (x, y) sequentially acquired at 0.05 μm intervals (z = 5 μm). Scale bars are 10 μm (a–c). The length indicates the scale of each unit (d,f,g). Hereafter, all images were reconstructed with 3D deconvolution. See also Supplementary Movie 1.